Murine
B16 melanoma expresses the
ganglioside GM3. GM3 shed from
tumor cells is immunosuppressive and promotes
tumor growth. Reduction or elimination of the shed GM3 could be therapeutic, and the anti-GM3
antibodies may reduce and clear the shed
ganglioside. To test this hypothesis, mice were challenged with
tumor cells, with or without inducing anti-GM3 antibody response. Since
gangliosides are poor immunogens and T-cell independent
antigens, an adjuvant (
monophosphoryl lipid A (MPL), a non-toxic
lipid A of Salmonella), directed against B-cells, was employed. MPL was incorporated onto
liposomes and into the surface membrane of B16 mouse
melanoma cells; both are rich in GM3. C57BL/6J mice immunized with MPL-
liposomes or MPL-B16 cells responded with elevated levels of anti-GM3
IgM. Non-immunized mice or mice immunized with B16 cells alone or
ganglioside GM3 alone (without MPL) elicited poor anti-GM3
IgM response, confirming the GM3's immunologic crypticity and MPL's immunopotentiating effect. MPL's immunopotentiating effect was improved by coupling it to
melanoma cell membranes. C57BL/6J mice were immunized with irradiated B16 alone or MPL alone or MPL-conjugated irradiated B16. After three weekly immunizations, each mouse received a challenge dose of viable syngeneic B16. Neither MPL alone nor B16 alone had a significant effect on
tumor growth or host survival; however, administration of MPL-conjugated B16 cells significantly prevented
tumor growth and prolonged survival. Our results indicate that MPL-incorporated B16 cells augment the anti-GM3
IgM response, which may reverse GM3-induced immunosuppression by eliminating
tumor-derived GM3, and restore immunocompetence.